目的 比较高脂饮食、高脂饮食联合腹腔注射链脲佐菌素（STZ）、plin1敲除（plin1-/-）和plin1敲除（plin1-/-）加高脂饮食对小鼠胰岛素抵抗（IR）的影响程度。方法 C57BL/6J小鼠分为5组：普通饮食组（W-N）、高脂饮食组（W-H）、高脂饮食+STZ组（W-S）、plin1-/-普通饮食组（K-N）和plin1-/-高脂饮食组（K-H）,每组8只,喂养12 w,测量小鼠体重,空腹血糖（FBG）,葡萄糖耐量（GTT）和胰岛素耐量（ITT）,并计算GTT和ITT曲线下面积,12 w后收集血清,处死小鼠,取出脂肪组织,检测空腹血清胰岛素水平（FINS）、血清甘油三酯（TG）、游离脂肪酸（FFA）,计算稳态模型评估胰岛素抵抗指数（HOMA-IR）、Western blotting检测脂肪组织磷脂酰肌醇3激酶（PI3K）、蛋白激酶B（AKT）、葡萄糖转运体4（GLUT4）蛋白表达水平。结果 与W-N组比较,其余四组小鼠FBG、GTT和ITT曲线下面积、HOMA-IR、血清TG均明显升高、PI3K、p-AKT和GLUT4蛋白表达均显著下调（P<0.05）,W-H组体重显著升高（P<0.05）,K-N组体重显著下降（P<0.05）,K-N组、K-H组、W-S组FINS显著升高（P<0.05）,W-H组FFA显著升高（P<0.05）,K-N组FFA显著降低（P<0.05）。与K-N组相比,W-H组、K-H组和W-S组小鼠体重显著升高（P<0.05）;W-H组小鼠血清FFA显著升高（P<0.05）;W-S组小鼠FBG显著升高,PI3K蛋白表达显著下调（P<0.05）;K-H组和W-S组GTT和ITT曲线下面积、HOMA-IR、血清TG、FFA显著升高、p-AKT和GLUT4蛋白表达显著下调（P<0.05）。W-H组小鼠FBG、GTT和ITT曲线下面积、FINS、HOMA-IR、TG、PI3K、p-AKT、GLUT4蛋白表达无明显变化。结论 plin1-/-诱导小鼠IR程度与高脂饮食喂养的小鼠相当,但低于高脂饮食+STZ诱导的小鼠,高脂饮食喂养的plin1-/-小鼠IR程度大于plin1-/-小鼠。
Abstract
Objective To compare the effects of high-fat diet, high-fat diet combined with intraperitoneal injection of streptozotocin (STZ), plin1 gene knockout (plin1-/-) and plin1 gene knockout (plin1-/-) plus high-fat diet on insulin resistance in mice. Methods C57BL/6J mice were divided into five groups: normal diet group (W-N), high-fat diet group (W-H), high-fat diet + STZ group (W-S), plin1-/-+ normal diet group (K-N), plin1-/-+ high-fat diet group (K-H). Each group had eight rats and was fed for 12 weeks. The body weight, fasting blood glucose (FBG), glucose tolerance (GTT) and insulin tolerance (ITT) were measured and the areas under the curve (AUC) of GTT and ITT were calculated. The mice were sacrificed after 12 weeks,and the serum and adipose tissue were collected. Fasting serum insulin (FINS), serum triglyceride (TG) and free fatty acids (FFA) were measured and homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. The protein expression levels of phosphatidylinositol 3 kinase (PI3K), protein kinase B(AKT) and glucose transporter 4(GLUT4) in adipose tissue were detected by Western blotting. Results Compared with the W-N group, the FBG, AUC of GTT and ITT, HOMA-IR, serum TG in the mice from other four groups were significantly increased, and the protein expressions of PI3K, p-AKT and GLUT4 were significantly decreased (P<0.05). The body weight of the W-H group was significantly increased (P<0.05), and that of the K-N group was significantly decreased (P<0.05). FINS was significantly elevated in the K-N, K-H and W-S groups (P<0.05), and FFA was significantly increased in the W-H group but reduced in the K-N group (P<0.05). Compared with the K-N group, the body weight of the W-H group, K-H group and W-S group was significantly increased (P<0.05). Serum FFA was significantly higher (P<0.05) in the W-H group. In the W-S group, FBG was significantly elevated and PI3K protein expression was dramatically downregulated (P<0.05). The AUC of GTT and ITT, HOMA-IR, serum TG and FFA in the K-H and W-S groups were significantly increased, and the protein expressions of p-AKT and GLUT4 were significantly decreased (P<0.05). There were no significant changes in FBG, AUC of GTT and ITT, FINS, HOMA-IR, TG, and the protein expressions of PI3K, p-AKT and GLUT4 in the W-H group. Conclusion The change of IR induced by plin1-/- is equivalent to that induced by high-fat diet, but lower than that induced by high-fat diet +STZ. The changes of IR in plin1-/- mice fed with the high-fat diet is higher than in plin1-/- mice fed the normal diet.
关键词
围脂滴蛋白 /
基因敲除 /
胰岛素抵抗
Key words
plin1 /
gene knockout /
insulin resistance
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基金
国家自然科学基金（No.201901D111182）
